This invention relates to a vaccine containing herpes virus for the prevention of atherosclerosis. More particularly the invention relates to a herpes vaccine containing several types of whole killed herpes viruses that affect humans and that acts as a prophylaxis against pathogenic development of atherosclerotic plaque in a mammalian subjected susceptible thereto.
It is generally accepted that atherogenesis is triggered by primary injury to the endothelial lining of the arterial walls. This injury is believed to be the result of exposure of the underlying smooth muscle tells to several factors of non-infectious origin (hormones, low density lipoproteins, growth factors, among others). The prevailing view is that human atherosclerosis (AS) is a pleiotropic process with various causes. See Ross, R., The Pathogenesis of Atherosclerosis: An Update, New England J. Med.,314, 488 to 500 (1986).
A fundamentally new etiological factor: herpes virus infection-was reported by Fabricant et al, who demonstrated that chickens infected with Marek Disease Virus (MDV), have an unusually high incidence of atherosclerotic plaque-(ASP) in the arteries. See Fabricant, C. G. et al, Virus-Induced Cholesterol Crystals, Science, 181, 566 to 567 (1973); and Fabricant, C. G. et al, Virus-Induced Atherosclerosis, J. Exp. Med., 148, 335 to 340 (1978). Since that time data have been accumulated suggesting the role of herpes virus in AS in humans. It was shown that different herpes viruses can alter smooth muscle cells lipid metabolism and induce cholesterol and cholesterol ester accumulation in these cells. See Fabricant, C. G. et al, Herpes Virus Infection Enhances Cholesterol and Cholesterol Ester Accumulation in Cultured Arterial Smooth Muscle Cells, Am. J. Pathol, 105, 176 to 184 (1981); Fabricant, C. G. et al, Herpes Virus-Induced Atherosclerosis in Chickens, Fed. Proc., 42, 2476 to 2479 (1983); Melnick, J. L. et al, Cytomegalovirus Antigen within Human Arterial Smooth Muscle Cells, Lancet, ii, 644 to 647 (1983); Gyorkey, F. et al, Herpesviridae in the Endothelial and Smooth Muscle Cells of Proximal Aorta in Atherosclerotic Patients, Exp. Mol. Pathol, 40, 328 to 339 (1984); Hajjar et al, Virus-Induced Atherosclerosis: Herpes Virus Infection Alters Aortic Cholesterol Metabolism and Accumulation, Am. J. Pathol., 122, 62 to 70 (1986); Adam et al, High Levels of Cytomegalovirus Antibody in Patients Requiring Vascular Surgery for Atherosclerosis, Lancet, 2, 291 to 293 (1987); Petrie, Association of Herpesvirus/Cytomegalovirus Infections with Human Atherosclerosis, Prog. Med. Virol., 35, 21 to 42 (1988); Grattan, M. T. et al, Cytomegalovirus Infection is Associated with Cardiac Allograft Rejection and Atherosclerosis, J. A. Med. Assoc., 261, 3561 to 3566 (1989); Mc Donald, K. et al, Association of Coronary Artery Disease in Cardiac Transplant Recipients with Cytomegalovirus Infection, Am. J. Cardiol., 64, 359 to 362 (1989); Visser et al, Granulocyte-Mediated Injury in Herpes Simplex Virus-Infected Human Endothelium, Lab. Invest., 60, 296 to 304 (1989); Melnick, J. L. et al, Possible Role of Cytomegalovirus in Atherogenesis, J. Am. Assoc., 263, 2204 to 2207 (1990); Bruggeman, C. A. et al, The Possible Role of Cytomegalovirus in Atherogenesis, Prog. Med. Virol., 38, 1 to 26 (1991); Melnick, J. L. et al, Accelerated Graft Atherosclerosis Following Cardiac Transplantation; Do Viruses Play a Role?, Clin. Cardiol., 14 (Supp. II), 21 to 26 (1991); and Hajjar , D. P., Viral Pathogenesis of Atherosclerosis, Am. J. Pathol., 133, 1195 to 1211 (1991).
In addition the DNA of various herpes viruses showed positive hybridization with ASP DNA; see Benditt, E. P. et al, Viruses in the Etiology of Atherosclerosis, Proc. Natl. Acad. Sci., 80, 6386 to 6389 (1983); Pyrzak, R. et al, Detection of Specific DNA Segments of Marek""s Disease Herpes Virus in Japanese Quail Susceptible to Atherosclerosis, Atherosclerosis, 68, 77 to 85 (1987); Petrie, B. L. et al, Nucleic Acid Sequences of Cytomegalovirus in Cultured Human Arterial Tissue, J. Inf. Dis., 155, 158 to 159 (1987); Yamashiroya, H. M. et al, Herpesviridae in Coronary Arteries and Aorta of Young Trauma Victims, Am. J. Pathol, 130, 71 to 79 (1988); and Hendrix, M. G. R. et al, The Presence of Cytomegalovirus Nucleic Acids in Arterial Walls of Patients Suffering From Grade III Atherosclerosis, Am. J. Pathol., 134, 1151 to 1157 (1989).
No systematic attempts to demonstrate a viral presence in ASP by direct isolation of infectious HSV from ASP and by detection of viral replication in ASP by Electron Microscopy have been reported. A viral presence in ASP would explain the presence of HSV-like DNA in ASP, and redirect research to determine the molecular mechanisms of viral involvement in etiology of atherosclerosis. In such a case, the possibility of a contamination of ASP in the blood vessels by HSV also has to be excluded.
None of the above references deals with the preparation of a vaccine against any form of the herpes virus infection. The following reference deals with the preparation of a herpes vaccine against Marek""s Disease Herpes-Virus in chickens: Fabricant, J. et al, Vaccination Prevents Atherosclerosis Induced by Marek""s disease Herpesvirus, College of Veterinary Medicine and Medicine, Cornell University, Ithaca and New York, N.Y. The reference appeared as an abstract in the Federation of American Societies for Experimental Biology, 65th Annual Meeting, Atlanta (1981).
The vaccine employed against Marek""s Disease Herpesvirus in chickens was derived from Turkey herpesvirus (HVT). There is no indication that a vaccine against atherosclerosis caused by human herpes virus could be prepared. There is no suggestion to employ a herpes vaccine containing whole killed viruses from many different types of herpes with the ability to serve as a prophylaxis to the pathogenic development of atherosclerotic plaque.
U.S. Pat. No. 4,038,381 is directed to a vaccine for the prevention and treatment of vascular conditions, comprising a combination of a tuberculosis antigen with an antiherpes vaccine. The reference indicates that neither component per se of the vaccine had any known utility in the prevention or treatment of vascular disease. Nor is there any suggestion in the reference to prepare a herpes vaccine containing whole killed viruses from many different types with the ability to serve as a prophylaxis to the pathogenic development of atherosclerotic plaque.
It is the object of the invention to provide a vaccine as a prophylaxis against pathogenic development of atherosclerotic plaque in a mammalian subject susceptible thereto.
We have discovered such a vaccine effective as a prophylaxis in a mammalian subject, including man, against the pathogenic development of human atherosclerotic plaque. The vaccine comprises a mixture of inactivated whole viruses of the following strains:
Herpes Simplex Virus 1;
Herpes Simplex Virus 2;
Herpes Simplex Virus 6;
Human Cytomegalovirus; and
Epstein-Barr Virus;
in combination with a pharmaceutically acceptable inert vaccine carrier such as normal saline or a physiological oil in an adequate concentration of said inactivated whole viruses. Preferably there is present 15 xcexcg per of each whole killed virus per ml of pharmaceutical composition. Thus the total amount of pure viruses in one dose of vaccine is 75 xcexcg.
Each of the five abovementioned whole killed viruses is preferably. present in the compositions in an amount of 15 to 25% by either weight or volume. More preferably equal amounts of each of the whole killed virus are present, that is the compositions contain about 20% by weight or volume of each of the five whole killed viruses as active ingredient.